OBJECTIVETo study the effects of Chinese medicine tetramethylpyrazine (TMP) on intracellular free calcium ([Ca2+]i) in cultured penis corpus cavernosum smooth muscle cell (PCSMC) in rabbits.

METHODSBy using laser scanning confocal microscope (LSCM), the [Ca2+]i fluorescence signal changes was investigated in cultured PCSMC loaded with Ca2+ indicator Fluo-3/AM and divided into potassium chloride(KCl) group and norepinephrine (NE) group. Compared with verapamil (Ver), the effects of TMP was observed in different concentrations on [Ca2+]i increase induced by high potassium and NE.

RESULTSTMP had no obvious effect on resting PCSMC [Ca2+]i. It was found that 1, 10, 100 mumol/L TMP significantly inhibited [Ca2+]i increase induced by high potassium-depolarization. The peak inhibition rates were (38.6 +/- 3.0)%, (44.1 +/- 2.4)% and (53.7 +/- 4.1)% respectively. TMP could also inhibit cytosolic calcium pool release induced by 1 mumol/L NE. The peak inhibition rates were (13.9 +/- 2.7)%, (21.2 +/- 1.9)% and (29.5 +/- 3.6)% respectively.

CONCLUSIONSTMP can inhibit rabbit PCSMC [Ca2+]i significantly by suppressing voltage-dependent calcium channel and cytosolic calcium pool release. The effect, similar to Ver, signifies the important mechanism of erectile dysfunction (ED) therapy.

Animals ; Calcium ; metabolism ; Cells, Cultured ; Dose-Response Relationship, Drug ; Male ; Muscle, Smooth, Vascular ; cytology ; metabolism ; Penis ; blood supply ; metabolism ; Pyrazines ; pharmacology ; Rabbits ; Vasodilator Agents ; pharmacology

Objective：This study was designed to determine the expression of ICAM-1 in colorectal cancer and its clinical significance. Methods：The expression of ICAM-1 in 55 samples of colorectal carcinoma and 8 samples of adenomas were invested using immunohistochemical staining. Results：There were 41 positive cases in 55 colorectal cancer samples. The positive rate of ICAM-1 in colorectal cancer was significantly higher than that of adenomas （ P<0.05） , ICAM-1 in colorectal cancer was significantly correlated with lymph node metastasis. However， there was no significantly difference in ICAM-1 expression between adenomas and colorectal cancer without lymph node metastasis. Conclusion： Over-expression of ICAM-1 was significantly related to lymph node metastasis.

OBJECTIVETo explore the expression and sequence of human MutS homologue 2 (hMSH2) during different stages of human bronchial epithelial (16HBE) cells induced by cadmium chloride (CdCl2).

METHODSReverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemical staining (SP method) were used to measure the hMSH2 mRNA and protein expression in 16HBE cells and its different passage cells treated by CdCl2 (the 5th, 15th, 35th passage, and neoplasm cells from nude mice's tumor tissue). hMSH2 exon 6, hMSH2 exon 7, hMSH2 exon 8, hMSH2 exon 9, hMSH2 exon 12 of the 16HBE cells and neoplasm cells from nude mice's tumor tissue were amplified by polymerase chain reactions (PCR). The amplified DNA strips were purified. Then the exons were detected by DNA analysis.

RESULTSDuring the passages of 16HBE cells treated with CdCl2, the expression of hMSH2 gene were decreased gradually. The hMSH2 gene mRNA and protein expression levels of the CdCl2 transformed 35th 16HBE cells and tumorigenic cells of nude mice significant decreased compared with non-transformed 16HBE cells (P < 0.01). In the tumorigenic cells of nude mice induced by CdCl2, there were thymine (T) deletion in 1st, 2nd and 7th site of hMSH2 exon 8, there were adenine (A) deletion in 20th and 182th site of hMSH2 exon 9, there were adenine (A) insertion in 241st site of hMSH2 exon 12. All the mutations were frame shift mutation.

CONCLUSIONThe expression decreased and the mutation of hMSH2 gene may be the possible carcinogenic mechanism for CdCl2.

Animals ; Bronchi ; cytology ; Cadmium Chloride ; toxicity ; Cell Line ; Cell Transformation, Neoplastic ; chemically induced ; genetics ; metabolism ; Epithelial Cells ; drug effects ; metabolism ; pathology ; Humans ; Mice ; Mice, Nude ; MutS Homolog 2 Protein ; genetics ; metabolism ; Mutation ; RNA, Messenger ; genetics

OBJECTIVETo explore safety, indications and advantages of mapping and ablation of arrhythmia in children guided by Carto and Ensite system.

METHODSGuided by Carto system, radiofrequency catheter ablation (RFCA) was performed on 8 pediatric patients with tachycardia whose mean age was (6.2 + or - 1.7) years, mean weight was (18.0 + or - 2.0) kg. Guided by Ensite system, RFCA was performed on 10 pediatric patients with arrhythmia, 8 of them were ablated guided by Ensite Array system: 6 cases with premature ventricular contractions (PVCs), 2 cases with right atrial tachycardia, their mean age was (11.3 + or - 1.2) years, and mean weight (40.0 + or - 5.0) kg. The other two cases with W-P-W syndrome were ablated guided by Ensite Navx system.

RESULTGuided by Carto system, 8 cases were successfully mapped and ablated: 6 cases had incision atrial tachycardia, 1 case had left atrial tachycardia and 1 case had right atrial tachycardia. In 1 case with incision atrial tachycardia the condition recurred after 3 months, and was ablated again successfully. Guided by Ensite Array system, 6 cases with PVCs (in 2 originating from the right ventricular inflow tract and in 4 originating from the right ventricular outflow tract) and 2 cases with right atrial tachycardia were successfully mapped and ablated, PVCs of the first 6 cases were reduced from (32 333 + or - 4509) 24 h to (0-4)/24 h after ablation. In 1 case with automatic atrial tachycardia, mapping could not be done by Ensite Array system, because P wave could not be identified from T wave. Single bolus of adenosine 20 mg was given within 30 s to let ventricles stop for 2 s (cardio-ventricular pacing standby) until T wave vanished, mapping and ablation were operated again successfully, but another atrial tachycardia occurred 1 day later. Guided by Ensite Navx system, 2 cases with W-P-W syndrome were successfully ablated, operation under X-rays lasted for 8 and 10 min. In none of the 9 patients the disease recurred after follow-up for 6 months.

CONCLUSIONCarto system is suitable for mapping and ablation in pediatric patients with continuous tachycardia, especially with incision atrial tachycardia; Ensite Array system fits children older than 10 years with right heart discontinuous arrhythmia; and Ensite NavX system can set up model and display endocardial anatomic structure quickly. Compared with two-dimensional mapping system, the three-dimensional mapping system (Carto and Ensite) can display the origin of arrhythmia and activation sequence clearly, decrease difficulty of operation efficiently and diminish operation time under X-ray.

Arrhythmias, Cardiac ; physiopathology ; surgery ; Catheter Ablation ; methods ; Child ; Child, Preschool ; Electrophysiologic Techniques, Cardiac ; methods ; Humans ; Imaging, Three-Dimensional ; Treatment Outcome

Liver cancer is a common malignant tumor in China and a major health concern. We aimed to estimate the liver cancer incidence and mortality in China in 2010 using liver cancer data from some Chinese cancer registries and provide reference for liver cancer prevention and treatment. We collected and evaluated the incidence and mortality data of liver cancer in 2010 from 145 cancer registries, which were included in the 2013 Chinese Cancer Registry Annual Report, calculated crude, standardized, and truncated incidences and mortalities, and estimated new liver cancer cases and deaths from liver cancer throughout China and in different regions in 2010 from Chinese practical population. The estimates of new liver cancer cases and deaths were 358,840 and 312,432, respectively, in China in 2010. The crude incidence, age-standardized rate by Chinese standard population (ASR China), and age-standardized rate by world standard population (ASR world) were 27.29/100,000, 21.35/100,000, and 20.87/100,000, respectively; the crude, ASR China, and ASR world mortalities were 23.76/100,000, 18.43/100,000, and 18.04/100,000, respectively. The incidence and mortality were the highest in western regions, higher in rural areas than in urban areas, and higher in males than in females. The age-specific incidence and mortality of liver cancer showed a rapid increase from age 30 and peaked at age 80-84 or 85+. Our results indicated that the 2010 incidence and mortality of liver cancer in China, especially in undeveloped rural areas and western regions, were among high levels worldwide. The strategy for liver cancer prevention and treatment should be strengthened.
China ; epidemiology ; Female ; Humans ; Incidence ; Liver Neoplasms ; epidemiology ; mortality ; Male ; Registries ; Rural Population ; Sex Distribution ; Urban Population

OBJECTIVETo explore dysfunction mechanism of rat alveolar type II (AT-II) injured by bleomycin (BLM).

METHODSSD rats were injected with a single intratracheal dose of bleomycin or control saline. On day 7, 14, and 28 following intratracheal bleomycin or saline instillation, animals were killed under overdose of 1.5% sodium pentobarbital (0.25 ml/100 g, i.p.) and bronchoalveolar lavage fluid (BALF) from the lung was tested for the activity of pulmonary surfactant (PS) by the Whihelmy Film Balance. Several concentrations of bleomycin stimulated the culture of rat AT-II cells, and surfactant protein (SP) A, B, and aquaporin-1 (AQP) mRNA were analyzed by fluorescent quantitative polymerase chain reaction (FQ-PCR).

RESULTSThe activity of PS and hypoxemia significantly decreased on day 7 and improved on day 14 and completely recovered to normal status on day 28. SP-A, B, and AQP-1 mRNA expression in BLM-stimulated group were significantly lower than those in the control group (P<0.001).

CONCLUSIONBLM-injured AT-II cells decrease the levels of SP-A, B, and AQP-1 mRNA and cause PS dysfunction, resulting in hypoxemia and pneumonedema.

Animals ; Aquaporin 1 ; biosynthesis ; genetics ; Bleomycin ; administration & dosage ; toxicity ; Cells, Cultured ; Dose-Response Relationship, Drug ; Epithelial Cells ; drug effects ; metabolism ; Hypoxia ; chemically induced ; metabolism ; pathology ; Male ; Pulmonary Alveoli ; cytology ; drug effects ; Pulmonary Surfactant-Associated Protein A ; biosynthesis ; genetics ; Pulmonary Surfactant-Associated Protein B ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Time Factors

OBJECTIVETo observe the morphological and molecular biological peculiarities of the experimental autoimmune prostatitis (EAP) rat model made by SC purified prostate protein twice with immune adjuvant.

METHODSMale rats were intradermally immunized with a saline extract of male rat prostate glands (RPG) in Freund's complete adjuvant (FCA) and Pertussis-Diphtheria-Tetanus vaccine 0.5 ml i.p. at the 0 and 30th day, and the concentrations of the extract were respectively 5 mg/ml, 10 mg/ml and 15 mg/ml. At the 45th day, the rats were sacrificed and the morphological and molecular biological changes of the prostate specimens were observed to determine the effective concentration of RPG for a successful model.

RESULTSThe expression of inflammation genes such as TNF-alpha, IL-1beta, IL-2 and iNOS obviously increased in the high-dosage model group; LM, EM and in situ hybridization revealed appearant chronic inflammation response, but this was not the case in the other two dosage groups.

CONCLUSION15 mg/ml RPG mixed with FCA (1:1) 1.0 ml SC with Pertussis-Diphtheria-Tetanus vaccine 0.5 ml i.p. was an effective dosage for the successful model in our experiment.

Animals ; Autoimmune Diseases ; immunology ; metabolism ; pathology ; Diphtheria-Tetanus-Pertussis Vaccine ; administration & dosage ; Disease Models, Animal ; Freund's Adjuvant ; administration & dosage ; Injections, Intraperitoneal ; Male ; Prostate ; metabolism ; pathology ; Prostatitis ; immunology ; metabolism ; pathology ; Proteins ; administration & dosage ; Rats ; Rats, Wistar

OBJECTIVESTo study the relaxation effects of tetrandrine on the corpus cavernosum tissue of rabbit in vitro.

METHODS1. The fluctuation of the dose-response relaxation curves for the contraction of KCl induced by tetrandrine was observed with isolated rabbit corpus cavernosum tissue. 2. Isolated strips of rabbit corpus cavernosum tissue were precontracted with 10 mumol/L phenylephrine(PE). Relaxation in response to cumulative doses of tetrandrine was determined in the absence and presence of nitric oxide synthase inhibitor (L-NNA) and soluble guanylate cyclase inhibitor (methylthioninium).

RESULTS1. The dose-response curves of KCl were shifted to the right nonparallelly, and the maximal responses were depressed to (73.0 +/- 3.8)% and (41.5 +/- 3.4)%, respectively, in the presence of tetrandrine(10 mumol/L, 30 mumol/L). 2. On rabbit cavernosal muscle stripes precontracted with PE(10 mumol/L), increasing concentrations of tetrandrine (1 mumol/L, 10 mumol/L, 30 mumol/L and 100 mumol/L) showed dose dependent relaxation [(6.0 +/- 1.4)%, (21.3 +/- 2.2)%, (47.4 +/- 3.3)%, and (68.1 +/- 3.6)%, P < 0.01]. However, in the meantime, it was found that these relaxation effects were not affected by the presence of L-NNA and methylthioninium (P > 0.05).

CONCLUSIONSTetrandrine was effective in relaxing rabbit corpus cavernosum tissue in vitro in a dose-dependent style. The mechanism might be related with its blocking effect on calcium channel, but not the NO-cGMP passage.

Alkaloids ; pharmacology ; Animals ; Benzylisoquinolines ; pharmacology ; Dose-Response Relationship, Drug ; In Vitro Techniques ; Male ; Muscle Relaxation ; drug effects ; Nitric Oxide ; physiology ; Penis ; drug effects ; physiology ; Phenylephrine ; pharmacology ; Potassium Chloride ; pharmacology ; Rabbits

OBJECTIVETo observe the effect of the combination of Wenxia Changfu Formula ([see text], WCF) with cisplatin (CDDP) on inhibiting non-small cell lung cancer (NSCLC) in vitro and In Vivo and explore its mechanism from its effect on cell cycle.

METHODSIn vitro, WCF-containing serum was prepared and the rhubarb b1, emodin, and aconitine were detected qualitatively by high-performance liquid chromatogram (HPLC). A549 cell lines were treated with blank control (dimethyl sulfoxide), normal serum, normal serum with CDDP (1.25, 2.5, and 5.0 μg/mL, respectively), WCF-containing serum plus different doses of CDDP (1.25, 2.5, and 5.0 μg/mL, respectively). The inhibitory effect was detected by 3-(4,5)-dimethylthiazo(-zy1)-3,5-diphenylterazolium bromide (MTT). The cell cycle was detected by flow cytometry. The protein and mRNA expressions of cyclin D1, proliferating cell nuclear antigen (PCNA), retinoblastoma (Rb), and p16 were observed with immunofluorescence and RT-PCR, respectively. In Vivo, nude mice xenograft model was established and grouped into the control, CDDP, WCF, and combination groups. The combination's inhibition of tumor growth and influence on the weight, spleen, and thymus gland were observed.

RESULTSThe inhibitory rate of the combination against A549 cell lines excelled the CDDP alone significantly (P <0.05); the combination showed a synergism inhibitory effect (Q=1.19). Compared with the monotherapy, the combination increased the cell percentage in G(0)/G(1) phase and decreased the cell percentage in S phase significantly (P <0.05); the protein and mRNA expressions of cyclin D1, PCNA, and Rb were significantly reduced; the protein and mRNA expressions of p16 were significantly enhanced. Compared with the monotherapy, the combination inhibited the tumor growth significantly In Vivo and reduced the weight of tumor (P <0.05); compared with the CDDP group, the spleen and thymus gland index of the combination group were enhanced significantly (P <0.05).

CONCLUSIONSThe combination of WCF with CDDP significantly inhibited the A549 cell lines proliferation in vitro and the growth of the tumor In Vivo; it inhibited effectively the atrophy of the immune organ caused by chemotherapy. The combination inhibited overproliferation of A549 cell lines by arresting the G(0) /G(1) phase of cell cycle and affecting the protein and mRNA expressions of cell cycle-related proteins, cyclin D1, etc.

Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; pharmacology ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; genetics ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chromatography, High Pressure Liquid ; Cisplatin ; pharmacology ; therapeutic use ; Drug Synergism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Flow Cytometry ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Lung Neoplasms ; drug therapy ; genetics ; pathology ; Male ; Neoplasm Proteins ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Wistar ; Spleen ; drug effects ; pathology ; Thymus Gland ; drug effects ; pathology ; Xenograft Model Antitumor Assays

OBJECTIVETo prepare and characterize polyelectrolyte multilayer film coated microbubbles for use as ultrasound contrast agent (UCA) and evaluate its effects in ultrasonic imaging on normal rabbit's liver parenchyma.

METHODSPerfluorocarbon (PFC)-containing microbubbles (ST68-PFC) were prepared by sonication based on surfactant (Span 60 and Tween 80). Subsequently, the resulting ST68-PFC microbubbles were coated using oppositely charged polyelectrolytes by microbubble-templated layer-by-layer self-assembly technique via electrostatic interaction. The enhancement effects in ultrasonic imaging on normal rabbit's liver parenchyma were assessed.

RESULTSThe obtained microbubbles exhibited a narrow size distribution. The polyelectrolytes were successfully assembled onto the surface of ST68-PFC microbubbles. In vivo experiment showed that polyelectrolyte multilayer film coated UCA effectively enhanced the imaging of rabbit's liver parenchyma.

CONCLUSIONSThe novel microbubbles UCA coated with polyelectrolyte multilayer, when enabled more function, has no obvious difference in enhancement effects compared with the pre-modified microbubbles. The polymers with chemically active groups (such as amino group and carboxyl group) can be used as the outermost layer for attachment of targeting ligands onto microbubbles, allowing selective targeting of the microbubbles to combine with desired sites.

Animals ; Contrast Media ; chemistry ; Electrolytes ; chemistry ; Fluorocarbons ; chemistry ; Liver ; diagnostic imaging ; Microbubbles ; Polymers ; chemistry ; Rabbits ; Surface Properties ; Surface-Active Agents ; chemistry ; Ultrasonics ; Ultrasonography